Molecular Engineering Powered Dual-Readout Point-of-Care Testing for Sensitive Detection of Escherichia coli O157:H7

Escherichia coli O157:H7 (E. coli O157:H7) has become one of the major threats to public health and food safety. However, the culture method as a gold standard for the detection of E. coli O157:H7 requires laborious operations and a long processing time. Herein, we developed a dual-readout aggregation-induced emission nanoparticle-based lateral flow immunoassay (LFIA) for sensitive detection of E. coli O157:H7 to achieve a qualitative and quantitative assay for satisfying the applications under varying scenarios. 2,3-Bis(4-(bis(4-(tert-butyl)phenyl)amino)phenyl)fumaronitrile (BAPF), an aggregation-induced emission luminogen, was designed to achieve a strong molar extinction coefficient (3.0 × 10⁴ M^-1 cm^-1) and high quantum yield (33.28%), which was further verified by a large rotation angle and low energy gap. Subsequently, BAPFs were integrated into a nanostructured system to form excellent water-soluble nanoparticles (BAPFNPs) for the detection of E. coli O157:H7 with colorimetric and fluorescent readout. The designed BAPFNPs-based LFIA (BAPFNPs-LFIA) exhibited nearly qualitative ability with gold nanoparticles-LFIA (AuNPs-LFIA) and a 9 times enhancement compared with quantum beads-LFIA (QBs-LFIA) in quantitative aspect. Especially, FL-BAPFNPs-LFIA could detect E. coli O157:H7 earlier than QBs-LFIA and AuNPs-LFIA when samples with low E. coli O157:H7 concentrations were cultured. Overall, the proposed strategy revealed that versatile BAPFNPs have great potential as reporters for dual-readout ability and enhancing detection sensitivity for rapid and accurate pathogenic bacteria assay.